Blood, Lymph, and Immunity:

Blood composition
Blood is a red fluid that is carried by the blood vessels of the circulatory system. It is composed
of a fluid part; the plasma, and a solid part; the blood cells (Fig. 1). Plasma forms part of
extracellular fluid (ECF). Each constituent of the blood plays a specific part in the overall
function of blood.

Fig. 1: Composition of blood

Plasma
Plasma is the liquid part of the blood that separates out when a blood sample is spun in a
centrifuge. The main constituent is water (about 90%) in which are a number of dissolved
substances being transported from one part of the body to another. These include carbon
dioxide in solution, nutrients such as amino acids, glucose and fatty acids, waste materials such
as urea, hormones, enzymes, antibodies and antigens.
In addition to these, plasma contains:
• Mineral salts: The main mineral salts found in ECF are sodium and chloride, but potassium,
calcium, magnesium and bicarbonate are also present. The functions of these mineral salts
include maintaining osmotic balance and maintaining pH by acting as buffers. Calcium has a
number of essential roles in the body, including blood clotting, muscle contraction and nerve
function.
• Plasma proteins help to maintain the osmotic pressure of the blood because they are too
large to pass out of the circulation. This has the effect of retaining fluid in the blood by osmosis;
in other words, it prevents too much water from ‘leaking’ out into the extracellular spaces. If
this did occur then the volume of the blood would decrease and the blood pressure would fall,
with serious consequences. The most important proteins are:
○ Albumin helps to maintain the osmotic concentration of the blood (i.e., holds the water in the
blood).
○ Fibrinogen and prothrombin are involved in the clottingmechanism of the blood.
○ Immunoglobulins are the antibodies produced by the immune system of the body.
Albumin, fibrinogen and prothrombin are produced by the liver, but the immunoglobulins are
produced by the cells of the immune system.
Blood cells
The blood cells make up the solid component of blood and can be divided into three types:
• Erythrocytes: the red blood cells
• Leucocytes: the white blood cells
• Thrombocytes: the platelets, which are cell fragments
Before studying the different types of blood cell it is useful to understand a number of terms.
• Haemopoiesis: the formation of all types of blood cell
• Erythropoiesis: the formation of red blood cells or erythrocytes
• Lymphoid tissue: found in the lymph nodes and spleen and produces agranular leucocytes
(i.e., lymphocytes and monocytes)
• Myeloid tissue: found in the red bone marrow and responsible for the formation of
erythrocytes and granular leucocytes (i.e., neutrophils, eosinophils and basophils)
• Serum: plasma minus the clotting factors fibrinogen and prothrombin; can be obtained by
allowing a blood sample to clot naturally
Red cells/ Erythrocytes
Also known as red blood cells or red blood corpuscles, erythrocytes are the most numerous
blood cell; there are about 6–8 million per cubic millilitre of blood (Fig. 2). Their function is to
transport oxygen and a small proportion of carbon dioxide around the body (most carbon
dioxide is carried in solution in the plasma).
Mature erythrocytes are biconcave circular discs about 7 μm in diameter. Erythrocytes contain a
red pigment called haemoglobin, which is a complex protein containing iron. They are the only
cells in the body without a nucleus, which allows a greater amount of haemoglobin to be
packed into a relatively small cell. Erythrocytes are surrounded by a thin, flexible cell
membrane, which enables them to squeeze through capillaries. Their shape and thin cell
membrane gives them a large surface area for gaseous exchange and allows oxygen to diffuse
across into the cell, where it combines with the haemoglobin to form oxyhaemoglobin.
Erythrocytes are formed from undifferentiated stem cells within the bone marrow by a process
known as erythropoiesis. The stem cells change into erythroblasts, which have a nucleus. The
cell begins to acquire haemoglobin and its nucleus shrinks; it is now known as a normoblast. As
the cell develops further it becomes a reticulocyte, at which point the nucleus consists only
of fine threads in the cytoplasm known as Howell–Jolly bodies. Eventually, the nucleus
disappears and the mature erythrocyte is released into the circulation. This process takes 4–7
days.
If there is a shortage of erythrocytes (e.g., acute haemorrhage or iron-deficiency anaemia),
reticulocytes are also released into the circulation to help make up the deficit. These can be
seen on a blood smear stained with methylene blue, which is a specific stain for reticulocytes.
A circulating erythrocyte has a lifespan of about 120 days, after which it is destroyed in the
spleen or lymph nodes. The iron from the haemoglobin is recycled back to the bone marrow
and the remainder is converted by the liver into the bile pigment bilirubin and excreted in bile.
The production of red blood cells is controlled by a hormone called erythropoietin, which is
released by cells in the kidney in response to low oxygen levels in the tissues. Erythropoietin
stimulates the stem cells in the bone marrow to produce more erythrocytes.
Chronic kidney disease can result in anaemia due to a decrease in the production of
erythropoietin, and this form of anaemia is classed as non-regenerative.

Fig. 2: The cellular components of blood.

White cells/ Leucocytes
Also known as white blood cells, leucocytes are much less numerous than red blood cells and
the cells contain nuclei. Leucocytes can be classified as either granulocytes or agranulocytes
depending upon whether they have visible granules in their cytoplasm when stained and
viewed under a microscope (Fig.2). The function of leucocytes is to defend the body against
infection.
Granulocytes
This type of leucocyte is produced within the bone marrow and makes up approximately 70% of
all leucocytes. They have granules within their cytoplasm and have a segmented or lobed
nucleus that can vary in shape. They are referred to as polymorphonucleocytes or PMNs
(meaning many-shaped nuclei). They can be further classified according to the type of stain they
take up: neutral, basic or acidic. There are three types of granulocytes:
• Neutrophils take up neutral dyes and the granules stain purple (Fig. 2). Immature neutrophils
have a nucleus that looks like a curved band and are known as band cells. Neutrophils are the
most abundant of the leucocytes, forming about 90% of all granulocytes. They are able to move
through the endothelial lining of the blood vessels into the surrounding tissues and engulf
invading bacteria and cell debris by phagocytosis, thus helping to fight disease. A neutrophilia or
raised numbers of neutrophils indicates the presence of an infective process, while a
neutropenia or lack of white cells may be characteristic of certain viral infections.
• Eosinophils take up acidic dye and the granules in their cytoplasm stain red (Fig. 2). They are
involved in the regulation of the allergic and inflammatory processes and secrete enzymes that
inactivate histamine. Eosinophils play a major role in controlling parasitic infestation. An
eosinophilia or raised numbers of eosinophils occurs in response to parasitic infestation.
• Basophils take up basic or alkaline dyes and the granules in the cytoplasm stain blue (Fig. 2).
Basophils secrete histamine, which increases inflammation, and heparin, which is a natural
anticoagulant preventing the formation of unnecessary blood clots. Basophils are present in
very small numbers in normal blood.
Agranulocytes
Agranulocytes have a clear cytoplasm. There are two types:
• Lymphocytes are the second most common type of white blood cell, forming 80% of all
agranulocytes (Fig. 2). Lymphocytes are the main cell type of the immune system and are
formed in lymphoid tissue, although they originate from stem cells in the bone marrow.
Lymphocytes are responsible for the specific immune response, and there are two different
types: the B lymphocytes, which produce antibodies and are involved in humoral immunity, and
the T lymphocytes, which are involved in the cellular immune response.
• Monocytes have a horseshoe-shaped nucleus and are the largest of the leucocytes, although
they are only present in small numbers (Fig.2). They are phagocytic cells and when they migrate
to the tissues they mature and become known as macrophages.
Lymphoma is a type of cancer that occurs when the B or T lymphocytes begin growing and
multiplying uncontrollably. The abnormal lymphocytes collect in one or more of the lymph
nodes or in lymphatic tissue (e.g., the spleen) and form a tumour.
Platelets/ Thrombocytes
Thrombocytes, or platelets, are cell fragments formed in the bone marrow from large cells
called megakaryocytes. They are small discs with no nuclei and are present in the blood in large
quantities. Platelets are involved in blood clotting.
Thrombocytopenia is the term used for a reduction in the number of platelets. Decreased
production may be linked to a bone marrow dysfunction, which may be as a result of a viral
infection, or if the animal is undergoing chemotherapy.
Blood clotting
The ability to form a blood clot is one of the most important defense mechanisms in the body. It
means that injured blood vessels can be sealed and excessive blood loss can be prevented.
Blood clotting is essential for wound healing and also prevents the entry of pathogenic
microorganisms into the wound. The formation of a blood clot is complicated and involves a
number of different chemical factors in the blood. It is described as a cascade mechanism
because one step leads on to another in a similar way to a cascade of water.
To simplify the process we will only consider the main steps.
When a blood vessel or a tissue is damaged the following happens:
1. Platelets stick to the damaged blood vessel and to one another to form a seal. The platelets
release an enzyme called thromboplastin.
2. In the presence of thromboplastin and calcium ions, the plasma protein prothrombin is
converted to the active enzyme thrombin. (Vitamin K is essential to the blood clotting
mechanism and is required for the manufacture of prothrombin in the liver.)
3. Thrombin then converts the soluble plasma protein fibrinogen into a meshwork of insoluble
fibres called fibrin. The presence of calcium is an essential factor.
4. The fibrin fibres form a network across the damaged area that traps blood cells and forms a
clot. This seals the vessel in what is often called a ‘scab’ and further blood loss from the wound
is prevented.
The normal clotting time in a healthy animal is 3–5 min but it may be affected by a number of
factors. It may be reduced by:
• Surface contact with materials that will act as foundation for the clot (e.g., gauze swabs).
• Raising the environmental temperature: keeping the animal in a warm kennel.
Clotting time may be increased by:
• Lack of vitamin K, which is needed by the liver to form prothrombin in the blood (seen in
warfarin poisoning, which interferes with levels of vitamin K).
• Liver disease: the liver manufactures the plasma proteins involved in clotting.
• Genetic factors (e.g., haemophilia), which affect the availability of some clotting factors.
• Systemic diseases such as anthrax, canine infectious hepatitis and viral haemorrhagic disease
all cause subcutaneous haemorrhages due to interference with blood clotting.
• Thrombocytopenia: Lack of platelets may be seen in some forms of leukaemia.
• Lack of blood calcium: This feature is used in the lab to prevent blood samples clotting.
Chemicals such as EDTA (ethylene diamine tetra-acetic acid), citrate and oxalate all combine
with calcium in the blood and prevent it being involved in the clotting process.
• Parasitic disease: Infestation with Angiostrongylus vasorum (lungworm) can cause clotting
dysfunction and subsequent haemorrhage (e.g., in the eye, which is described as hyphaema).
The body has its own natural anticoagulant called heparin. This is secreted by the basophils and
prevents unwanted clots forming in the blood vessels and organs. If part of a clot, referred to as
an embolus, detaches, it may be carried around the body and block any of the vital blood
vessels, killing the animal.
Heparin is added to a saline solution to make heparinized saline. This is used to flush
intravenous catheters on their placement and to ensure their ongoing patency.
Lymphatics
The lymphatic system is part of the circulatory system of the body and is responsible for
returning the excess tissue fluid that has leaked out of the capillaries to the circulating blood.
The fluid within the system is called lymph and is similar to plasma but without the larger
plasma proteins. However, lymph contains more lymphocytes than are present in the blood.
The lymphatic system is composed of both lymphatic vessels and lymphoid tissue, which are
found in all regions of the body with a few exceptions such as the central nervous system and
bone marrow.
The functions of the lymphatic system are:
• To return excess tissue fluid that has leaked out of the capillaries to the circulating blood
• To remove bacteria and other foreign particles from the lymph in specialised filtering stations
known as lymph nodes
• To produce lymphocytes, which produce antibodies; the lymphatic system may also be
considered as part of the immune system
• To transport the products of fat digestion and the fat-soluble vitamins from the lacteals of the
intestinal villi to the circulation
• Absorbs fatty acids
The lymphatic system consists of the following parts:
• Lymphatic capillaries
• Lymphatic vessels
• Lymphatic ducts
• Lymph nodes
• Lymphatic tissues
Lymphatic capillaries
Excess tissue fluid is collected up by the smallest of the lymphatic vessels, the lymphatic
capillaries. These are thin-walled, delicate tubes, which form networks within the tissues. In the
villi of the small intestine the lymph capillaries are called lacteals and are responsible for
collecting up the products of fat digestion.
Lymphatic vessels
The lymphatic capillaries merge to form the larger lymphatic vessels, which have a similar
structure to veins and possess numerous closely spaced valves. Lymph flow is mainly passive
and relies on the contraction of the surrounding muscles to move the lymph along. The valves
prevent backflow and pooling of lymph in the vessels.
Lymphatic ducts
The lymphatic vessels enter the larger lymphatic ducts (Fig. 3), which drain lymph into blood
vessels leading to the heart and so return it to the circulation. The major lymphatic ducts are:
• The right lymphatic duct is the smaller of the two major ducts and drains lymph from the right
side of the head, neck and thorax, and right forelimb. It empties into the right side of the heart
via either the right jugular vein or cranial vena cava.
• The thoracic duct is the main lymphatic duct and collects blood from the rest of the body. It
arises in the abdomen, where it is called the cisterna chyli, and receives lymph from the
abdomen, pelvis and hind limbs. As it passes through the aortic hiatus of the diaphragm and
enters the thorax, it becomes known as the thoracic duct and receives lymph from the left side
of the upper body and left forelimb. The thoracic duct empties into either the jugular vein or
cranial vena cava, near the heart.
• There is also a pair of tracheal ducts that drain the head and neck and empty either into the
thoracic duct or one of the large veins near the heart.
Fig. 3: Location of the lymphatic vessels and lymph nodes.
Chylothorax occurs when there is a leakage of lymphatic fluid (chyle) into the thoracic cavity.
This can be due to the rupture of the thoracic duct (e.g., from trauma, tumour, or surgery) and
the accumulation of fluid between the lungs and the pleura causes respiratory distress.
Lymph nodes
The lymph nodes (Fig. 4) are masses of lymphoid tissue situated at intervals along the lymphatic
vessels (Fig. 3). They are bean-shaped and have an indented region, called the hilus, where the
lymph vessels leave the node. A number of afferent lymphatic vessels carry lymph to each
lymph node and enter the node on its convex surface. The lymph leaves the node at
the hilus in the single efferent vessel.
Fig. 4: Structure of a lymph node.

Each node is surrounded by a fibrous connective tissue capsule. Inside, a network of tissue,
called trabeculae, extends from the capsule and provides support for the entire node. The tissue
of the node is divided into cortical and medullary regions. The cortex contains the germinal
centres or lymph nodules that produce the lymphocytes, which play an important part in the
immune system. The medulla is comprised of a reticular framework containing many phagocytic
cells. Lymph flows through spaces or sinuses within the tissue of the node.
All lymph must pass through at least one lymph node before being returned to the circulation.
Each node acts as a mechanical filter, trapping particles such as bacteria within the meshwork of
tissue. The particles are then destroyed by phagocytic cells. Lymph nodes are distributed
throughout the body and range in size. They can become enlarged during infection and are an
indication of a disease process in the drainage region. In a generalised infection or disease all
the lymph nodes may be enlarged; this is described as lymphadenopathy.
Some lymph nodes are superficial and can be palpated (Fig. 5).
These include:
• Submandibular nodes: two to five nodes in a group, lying at the edge of the angle of the jaw
• Parotid node: lies just caudal to the temporomandibular joint of the jaw
• Superficial cervical nodes also called the prescapular nodes: two on each side lying just in
front of the shoulder joint, at the base of the neck on the cranial edge of the scapula
• Superficial inguinal nodes: two nodes on each side lying in the groin, between the thigh and
the abdominal wall, dorsal to the mammary gland or penis
• Popliteal node: lies within the tissue of the gastrocnemius muscle, caudal to the stifle joint.
Lymphatic tissues
These include organs that contain lymphoid tissue and play an important part in the body’s
defense system.
Spleen
This is the largest of the lymphoid organs. The spleen is found within the greater omentum,
closely attached to the greater curvature of the stomach. It is a dark red haemopoietic organ
that is not essential for life, and therefore can be surgically removed if necessary, for example, if
ruptured or if a tumour develops. The spleen has a number of functions:
1. Storage of blood: It acts as a reservoir for red blood cells and platelets.
2. Destruction of worn-out red blood cells: The phagocytic cells engulf and destroy the
erythrocytes and preserve their iron content for re-use in haemoglobin synthesis.
3. Removal of particulate matter from the circulation: It ‘filters’ out foreign particles and
bacteria and the phagocytic cells destroy them.
4. Production of lymphocytes: It produces lymphocytes for use in the immune system.
Thymus
This is of the greatest importance in the young animal. The thymus lies in the cranial thoracic
inlet and cranial part of the thorax. It is active in late fetal and early postnatal life and is
responsible for the production of T lymphocytes that give rise to the cell-mediated immune
response. It begins to regress at the time of puberty and may eventually almost disappear.
Tonsils
These form a ring of lymphoid tissue around the junction of the pharynx with the oral cavity.
They can be seen, especially when reddened and enlarged, on either side of the pharynx at the
root of the tongue. They act as a ‘first line’ defense against microorganisms that enter the
mouth.
Gut-associated lymph tissue
is a component of the mucosa-associated lymphoid tissue (MALT) which works in the immune
system to protect the body from invasion in the gut. Owing to its physiological function in food
absorption, the mucosal surface is thin and acts as a permeable barrier to the interior of the
body. Equally, its fragility and permeability creates vulnerability to infection and, in fact, the vast
majority of the infectious agents invading the body use this route.
The functional importance of GALT in body's defense relies on its large population of plasma
cells, which are antibody producers, whose number exceeds the number of plasma cells in
spleen, lymph nodes and bone marrow combined. GALT makes up about 70% of the immune
system by weight; compromised GALT may significantly affect the strength of the immune
system as a whole.
The gut-associated lymphoid tissue lies throughout the intestine. In order to increase the
surface area for absorption, the intestinal mucosa is made up of finger-like projections (villi),
covered by a monolayer of epithelial cells, which separates the GALT from the lumen intestine
and its contents. These epithelial cells are covered by a layer of glycocalyx on their luminal
surface so as to protect cells from the acid pH.
New epithelial cells derived from stem cells are constantly produced on the bottom of the
intestinal glands, regenerating the epithelium (epithelial cell turnover time is less than one
week). Although in these crypts conventional enterocytes are the dominant type of cells, Paneth
cells can also be found. These are located at the bottom of the crypts and release a number of
antibacterial substances, among them lysozyme, and are thought to be involved in the control
of infections.
Underneath them, there is an underlying layer of loose connective tissue called lamina propria.
There is also lymphatic circulation through the tissue connected to the mesenteric lymph nodes.
Both GALT and mesenteric lymph nodes are sites where the immune response is started due to
the presence of immune cells through the epithelial cells and the lamina propria.
The GALT also includes the Peyer's patches of the small intestine and isolated lymphoid follicles
present throughout the intestine.
Immune system and Functions
The body has a number of natural defense systems such as the phagocytic blood cells that
engulf and destroy invading bacteria, the inflammatory response, and the ability of the blood to
form a clot, thus preventing blood loss and repairing damage. In addition to the phagocytic cells
in the blood, there is a system of phagocytic cells distributed around the body in the tissues
known as the reticuloendothelial system. These phagocytic cells are macrophages (agranular
leucocytes) and are referred to by a number of names depending on where they are found; in
connective tissue, for example, they are called histiocytes.
These defense mechanisms are all non-specific and the response is the same no matter what
the stimulus, such as wounding, trauma or invading pathogen. However, the body also has a
sophisticated immune system that provides a more specific response to a particular pathogen.

Reactions
The main type of cell in the immune system is the lymphocyte and there are two types of
specific immune response:
• Humoral (antibody-mediated) immune response involves the production of antibodies or
immunoglobulins by the B lymphocytes. When a particular type of antigen (a foreign substance
or invading organism that stimulates an immune response) enters the body, it stimulates the B
lymphocytes to produce a corresponding protein called an antibody, which combines with the
antigen and ‘neutralizes’ it.
• Cell-mediated immune response involves the use of T lymphocytes, which recognize and help
in the destruction of non-self-cells (i.e., cells that do not belong to the body), or any of the
body’s own cells that have been altered by virus infections.
Immunization
Vaccination is the introduction of antigenic material, such as a viral or bacterial element in an
inactivated or harmless form, which will stimulate the body’s immune response and prevent
infection by developing an immunity to that specific pathogen.
Passive
Passive immunization involves the production of antibodies in one animal by active
immunization and transfer to another. The donor animal can be bled and its serum administered
to susceptible animals to confer immediate but short-lived protection. The transfer of maternal
antibody to offspring via the placenta or colostrum is the natural (and very important) form of
passive immunization. Immune globulins may be produced in cattle against anthrax, in dogs
against distemper virus, and in cats against panleukopenia virus. Their most important role is in
protection against toxigenic organisms, such as tetanus, botulism, or Clostridium perfringens. It
can also be used in the treatment of snake envenomation. These immune globulins are
generally produced in young horses by a series of immunizing inoculations.
Tetanus immune globulin is given to animals to confer immediate protection against tetanus. At
least 1,500–3,000 IU of immune globulin should be given to horses and cattle; at least 500 IU to
calves, sheep, goats, and pigs; and at least 250 IU to dogs. The exact amount varies with the
amount of tissue damage, degree of wound contamination, and time elapsed since injury.
Tetanus immune globulin is of little use once clinical signs appear, although massive doses of up
to 300,000 IU may help.
Active
Active immunization involves administration of vaccines containing antigenic molecules (or
genes for these molecules) derived from infectious agents. In response, the animals mount
adaptive immune responses and develop prolonged, strong immunity to those agents. Vaccines
are by far the most effective way of controlling infectious diseases and, as a result, have
increased human and companion animal longevity and made intensive livestock production
possible.
Several criteria determine whether a vaccine can or should be used. First, the actual cause of
the disease must be determined. Although this seems self-evident, it has not always been
followed in practice. For example, although Mannheimia haemolytica can be isolated
consistently from the lungs of cattle with respiratory disease, these bacteria are not the sole
cause of this syndrome, and vaccines against the primary viral pathogens are required for full
protection. In some important viral diseases, such as equine infectious anemia, feline infectious
peritonitis, and Aleutian disease in mink, antibodies may contribute to the disease process, and
vaccination may therefore increase disease severity.
An ideal vaccine for active immunization should confer prolonged, strong immunity in
vaccinated animals and induce rapid onset of immunity. Ideally, depending upon the pathogen,
it should induce the most effective response, such as type 1 or type 2 immunity depending
upon the nature of the pathogen. It should preferably stimulate responses distinguishable from
those due to natural infection so that vaccination and eradication may proceed simultaneously.
Vaccination is not always an innocuous procedure; adverse effects can and do occur. Therefore,
all vaccinations must be governed by the principle of informed consent. The risks of vaccination
must not exceed those caused by the disease itself.